From owner-biotechniques@net.bio.net Wed Jan 03 22:00:00 1996 Path: biosci!biosci!not-for-mail From: cmody@acs.ucalgary.ca Newsgroups: bionet.journals.letters.biotechniques Subject: Spectrophotometric lipid concentration assay Date: 4 Jan 1996 07:02:51 -0800 Organization: The University of Calgary Lines: 10 Sender: daemon@net.bio.net Approved: MMCCARTHY@BIOTECHNET.COM Distribution: world Message-ID: NNTP-Posting-Host: net.bio.net Our objective is to determine the lipid concentration in yeast (cryptococcal) cell fractions acquired from glass bead mill homogenization. Concentrations of DNA, protein, and carbohydrate have already been determined spectrophotometrically. In order to complete the characterization of the aforementioned fractions lipid concentrations must be assayed. Any replies with recommendations of an inexpensive, non-radioactive lipid assay would be much appreciated. Thank you for your time. tfbruno@acs.ucalgary.ca From owner-biotechniques@net.bio.net Thu Jan 11 22:00:00 1996 Path: biosci!biosci!not-for-mail From: MMCCARTHY@biotechnet.com Newsgroups: bionet.journals.letters.biotechniques Subject: BioTechniques Database on the Web Date: 12 Jan 1996 06:25:09 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 12 Sender: daemon@net.bio.net Approved: MMCCARTHY@BIOTECHNET.COM Distribution: world Message-ID: <01HZX1VD08TU99H8EB@delphi.com> NNTP-Posting-Host: net.bio.net The BioTechniques database is now available on the BioTechniques home page at http://www.BioTechniques.com Restricted and inclusive searches can be conducted by title, author, keyword, abstract and "all text". Check it out! The site is still under construction and suggestions for improvements are welcome. Mary McCarthy Associate Editor, BioTechniques Moderator, bionet.journals.letters.biotechniques From owner-biotechniques@net.bio.net Thu Jan 11 22:00:00 1996 Path: biosci!biosci!not-for-mail From: MMCCARTHY@biotechnet.com Newsgroups: bionet.journals.letters.biotechniques Subject: BioTechniques 20(2), February 1996 Date: 12 Jan 1996 10:34:35 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 85 Sender: daemon@net.bio.net Approved: MMCCARTHY@BIOTECHNET.COM Distribution: world Message-ID: <01HZXALITNHE9BYS78@delphi.com> NNTP-Posting-Host: net.bio.net BioTechniques 20(2), February 1996 Table of Contents BENCHMARKS Combined protocol for selection and isolation of recombinant plasmids using mutliwell plates -Brady, J.P. Simplified probe preparation facilitates S1 nuclease analysis -Noti, J.D. and Reinemann, B.C. DNA fingerprinting of mammalian cell lines using nonradioactive arbitrarily primed PCR (AP-PCR) -Schlegel, J., Vogt, T., M=81nkel, K. and R=81schoff, J.=20 Differential display assay and analysis -Shoham, N.G., Arad, T., Rosin-Abersfeld, R., Mashiah, P., Gazit, A. and Yaniv, A.=20 Direct cloning of PCR products amplified with Pwo DNA polymerase -Hinnisdaels, S., Del-Favero, J. and Vauterin, M.=20 Creation of deletion, insertion and substitution mutations using a single pair of primers and PCR -Hughes, M.J.G. and Andrews, D.W. Preparation of magnetic oligo(dT) particles -Kolarova, H. and Hengerer, B.=20 Isolation of 3.5-kb fragments on magnetic solid supports -Kotsopoulos, S.K. and Shuber, A.P.=20 THE INTERNET ON-RAMP (Cyberspace for Biologists) SHORT TECHNICAL REPORTS Specific suppression of false-positive signals in the product-enhanced reverse transcriptase assay -Lugert, R., K=94nig, H., Kurth, R. and T=94njes, R.R. Evaluation of different amplification protocols for use in primer-extension preamplification -Casas, E. and Kirkpatrick, B.W. Rapid bright-field detection of oligonucleotide primed in situ (PRINS)-labeled DNA in chromosome preparations and frozen tissue sect= ions -Speel, E.J.M., Lawson, D., Ramaeckers, F.C.S., Gosden, J.R. and Hopman, A.H.N. Winner 1995 BioTechniques Purchasing Survey Sweepstakes BIOFEEDBACK RESEARCH REPORTS Cycling probe technology with RNase H attached to an oligonucleotide -Bekkaoui, F., Poisson, I., Crosby, W., Cloney, L. and Duck, P= . Rapid quantification of gene expression by competitive RT-PCR and ion-pair reversed-phase HPLC -Hayward-Lester, A., Oefner, P.J. and Doris, P.A. Detection and quantitation of unlabeled nucleic acids in polyacrylamide gels -Hendry, P. and Hannan, G. General approach to analysis of polymorphic short tandem repeat loci -Sprecher, C.J., Puers, C., Lins, A.M. and Schumm, J.W. Method for multiple portal vein infusions in mice: quantitation of adenovirus-mediated hepatic gene transfer -Vrancken Peeters, M.-J.T.F.D., Lieber, A., Perkins, J. and Kay, M.A.=20 Detection of protease activity using a fluorescence-enhancement globular substrate -Voss, E.W.,Jr., Workman, C.J. and Mummert, M.E. =20 PRODUCT APPLICATION FOCUS Precision 96-channel dispenser for microchemical techniques -Stanchfield, J., Wright, D., Hsu, S., Lamsa, M. and Robbins, A. Synthesis and purification in a single column on a high-throughput automated oligonucleotide production system -Baier, J., Kaufman, J., Mason, G., Wang, J., Wright, P., Andrus, A.=20 AD HOC REVIEWERS NEW PRODUCTS RECRUITMENT INDEX TO ADVERTISERS From owner-biotechniques@net.bio.net Thu Jan 11 22:00:00 1996 Path: biosci!biosci!not-for-mail From: MMCCARTHY@biotechnet.com Newsgroups: bionet.journals.letters.biotechniques Subject: BioTechniques 20(1), January 1996 Date: 12 Jan 1996 06:30:54 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 92 Sender: daemon@net.bio.net Approved: MMCCARTHY@BIOTECHNET.COM Distribution: world Message-ID: <01HZX1YG7QZY99H8EB@delphi.com> NNTP-Posting-Host: net.bio.net BioTechniques 20(1), January 1996 Table of Contents BENCHMARKS Device for the simultaneous screening of bacteriophage lambda clones -Espinosa, L. and Navarro, E. Preparation of T-overhang vectors with high PCR product cloning efficiency -Hadjeb, N. and Berkowitz, G.A. Simple 5' extension method allowing transcript discrimination -Breda, C., El Turk, J. and Esnault, R. Transfer technique for minimizing waste of sonified adjuvant emulsions -Spack, E.G. and Toavs, D. Removal of albumin from multiple human serum samples -Rengarajan, K., de Smet, M.D. and Wiggert, B. Measurement of 8-OHdG in DNA by HPLC/ECD: The importance of DNA purity -Laws, G.M. and Adams, S.P. Transient gene transfer into myotubes following differentiation in culture -Robey, R.B., Osawa, H., Printz, R.L. and Granner, D.K. Preparation of electro-competent E. coli using salt-free growth medium -Sharma, R.C. and Schimke, R.T. Rapid PCR method for site-directed mutagenesis on double-stranded plasmid DNA -Xu, X., Kang, S.-H., Heidenreich, O., Li, Q. and Nerenberg,M. Measurement of human growth hormone using a chemiluminescence assay and a "glow" luminometer -Guidon, P.T.,Jr., Peter, R. and Kumar, E. Simple method for adapting DNA fragments and PCR products to all of the commonly used restriction sites -Tsang, T.C., Harris, D.T., Akporiaye, E.T., Schluter, S.F., Bowden, G.T. and Hersh, E.M. Chemiluminescent detection of unique sequences on chromosomes after on-slide PCR -Xie, H. and Troyer, D.L. PCR-TIES (third irrelevant enzyme site)-mediated gene fusion method -Zeng, M. and Lapeyre, J.-N. NEW FEATURE: The Internet On-Ramp (Cyberspace for Biologists) SHORT TECHNICAL REPORTS Cloning strategies using a third irrelevant enzyme site (TIES) to overcome certain cloning problems -Zeng, M., Huang, B.-R. and Lapeyre, J.-N. Cloning of size-selected human immunoglobulin heavy chain rearrangements from third complementarity-determining region fingerprint profiles -Raaphorst, F.M., Tami, J. and Sanz, I.E. Gene transfer into subcultured endometrial cells using lipofection -Lascombe, I., Mougin, P., Vuillermoz, C., Adessi, G.L. and Jouvenot, M. Harvest protocol to reduce variability of soluble enzyme yield from cultured cells -Buskin, J.N., Gregory, D.L., LaFramboise, W.A. and Hauschka, S.D. Bicistronic vector for the creation of stable mammalian cell lines that predisposes all antibiotic-resistant cells to express recombinant protein -Rees, S., Coote, J., Stables, J., Goodson, S., Harris, S. and Lee, M.G. Enhanced cytochemical detection of viral proteins and RNAs using double-sided labeling and light microscopy. -Ding, X.S., Carter, S.A. and Nelson, R.S. RESEARCH REPORTS Expression of a bifunctional chimeric protein A-Vargula hilgendorfii luciferase in mammalian cells -Maeda, Y., Ueda, H., Hara, T., Kazami, J., Kawano, G., Suzuki, E., Nagamune, T. Synthesis of a new substrate for detection of lacZ gene expression in live Drosophila embryos -Minden, J.S. Construction of a restriction map and gene map of the lettuce chloroplast small single-copy region using Southern cross-hybridization -Mitchelson, K.R. PRODUCT APPLICATION FOCUS Vectors for expression and secretion of FLAG epitope-tagged proteins in mammalian cells -Chubet, R.G. and Brizzard, B.L. NEW PRODUCTS INDEX TO ADVERTISERS From owner-biotechniques@net.bio.net Mon Jan 22 22:00:00 1996 Path: biosci!biosci!not-for-mail From: BIOSCI Administrator Newsgroups: bionet.journals.letters.biotechniques Subject: BIOSCI miniFAQ, ver. 14-DEC-95 Date: 23 Jan 1996 06:55:27 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 199 Sender: daemon@net.bio.net Approved: MMCCARTHY@BIOTECHNET.COM Distribution: world Message-ID: <199601201000.CAA11185@net.bio.net> NNTP-Posting-Host: net.bio.net (LAST REVISION: 14-DEC-95) This BIOSCI "miniFAQ" is designed to answer the questions that come up the *most frequently*. The main BIOSCI FAQ (Frequently Asked Questions) is accessible on the World Wide Web at URL http://www.bio.net/. Contents: -------- 1) Using the WWW to access the BIOSCI/bionet newsgroups. 2) What to do about "spams," i.e., junk mail, ads, etc. 3) Examples of subscribing and unsubscribing to the mailing lists. 4) The BIOSCI user address and research interest directory. 1) Using the WWW to access the BIOSCI/bionet newsgroups. -------------------------------------------------------- As of 10 December 1995, all BIOSCI/bionet full newsgroups are accessible through the World Wide Web (WWW) at URL http://www.bio.net. One can read and reply publicly or privately to both recent postings and archived messages through one's Web browser if it is configured properly to send e-mail. Each newsgroup is equipped with its own WAIS index in addition to the master index for the entire set. The main BIOSCI home page also has access to the BIO-JOURNALS Table of Contents database WAIS index and the BIOSCI user address database described in another item further below. 2) What to do about "spams," i.e., junk mail, ads, etc. ------------------------------------------------------- BIOSCI is a set of parallel USENET newsgroups (the "bionet" groups), mailing lists, and a hypermail archive at URL http://www.bio.net/. The same postings are distributed on all media (except for a small number of mailing-list-only groups at net.bio.net). Unfortunately it is becoming a despicable practice on the Internet (by a few people out to make a fast buck) to do automated mass postings to thousands of newsgroups and mailing lists. These attempts to grab free advertising are refered to as "spams" in the usual, somewhat boneheaded, net terminology. USENET is more susceptible to this practice, and many spams originate on the USENET groups and then are passed on to the mailing lists. However, spammers also get lists of mailing addresses and hit these too, so neither medium is immune. What should you do personally if you get junk mail? --------------------------------------------------- Just delete it and move on without reading it further. Filing a protest is becoming increasingly useless because spammers are often disguising the addresses where the messages are sent from. Unless you really understand Internet mail systems, your attempt at protest by sending replies to the message will often end up being sent to the address of an innocent person that the spammer is victimizing. What can BIOSCI/bionet do to protect its newsgroups? ---------------------------------------------------- The only solution currently available is to moderate the newsgroup. If this newsgroup is already moderated, then you are in good shape. Moderation protects the USENET distribution from about 95% of the spams that are being sent to date and protects the mailing lists completely. Moderation means, however, that someone has to take the time to review each message before it goes out. We have set up software here that simply allows the moderator to forward to an address at net.bio.net messages that (s)he wishes to have distributed. This takes no more time than that needed to read the message and pass it on, say about 1 min. per message. Most newsgroups currently have a discussion leader who is responsible for their newsgroup. The discussions leaders and their e-mail addresses are listed in the BIOSCI Information Sheet which is available on the Web at http://www.bio.net/. If a newsgroup is being hit with too many junk postings, please contact the discussion leader for that group and see if there is interest in moderating the group. Please do not assume that by simply posting a complaint to the newsgroup itself, anyone on the BIOSCI staff will act on your complaint. With close to 100 newsgroups to run, the BIOSCI staff has to rely on the discussion leaders of each newsgroup to report problems directly to us at biosci-help@net.bio.net. We will moderate any of our newsgroups if the discussion leader tells us that the readership of the group wishes to do so and if a moderator is willing to do the work. For most BIOSCI/bionet groups, this entails only a few minutes of work each day. Moderating a newsgroup will resolve probably 95% of the junk postings on the USENET distribution. Unfortunately there are easy ways for determined spammers to override the moderation mechanism on USENET, but we can protect our e-mail subscribers from unwanted postings if the newsgroup is moderated. You can also access our newsgroups over the WWW at URL http://www.bio.net. While this Web interface will not stop spammers from trying to post to the groups, this will give you yet another way, besides using USENET news, to keep the junk out of your personal mail files. For those of you with local USENET news systems, the Web interface will also give you faster access to new newsgroups and recent postings. 3) Examples of subscribing and unsubscribing to the mailing lists. ------------------------------------------------------------------ PLEASE NOTE: The BIOSCI management does NOT act on subscription/unsubscription requests that are posted improperly to the newsgroups and mailing lists. People who do this only bother everyone on the lists to no avail. Please be sure to follow the proper procedures below. Gory details are in the BIOSCI Information sheets on the Web at http://www.bio.net. Below we give an example utilizing the METHODS-AND-REAGENTS list at both of our two BIOSCI sites: Users in the Americas and Pacific Rim countries who use the BIOSCI ------------------------------------------------------------------ node at computer net.bio.net: ---------------------------- A) Determine the "listname" which is the <=8 character mail address ^^^^^^^^^^^^^ for the group. These can be found in the BIOSCI Info. Sheet. For the METHODS-AND-REAGENTS group the mailing address is methods@net.bio.net. The listname is the portion of the address to the left of the @ sign, i.e., "methods". The listname is used with the "subscribe" and "unsubscribe" commands illustrated below. B) Mail all commands in the body of a mail message addressed to biosci-server@net.bio.net. Do NOT send commands to the newsgroup posting addresses! Leave the Subject: line blank, any text on it will be ignored. C) In the body of your message put one or more of the following commands with an "end" command on the last line, e.g., subscribe methods unsubscribe methods end Do NOT put your e-mail address or other text on these lines. The server only allows you to cancel your subscription if the address on your mail header matches the address on our mailing list. Please ask for help at biosci-help@net.bio.net if your address has changed, e.g., if you know you are on the list but the server tells you that you are not a member. Users in Europe, Africa, and Central Asia who use the BIOSCI node at -------------------------------------------------------------------- computer daresbury.ac.uk (also known as dl.ac.uk): ------------------------------------------------- To subscribe and unsubscribe to/from the BIOSCI lists, you need to specify the full USENET newsgroup name with "bionet-news." prepended. The USENET newsgroup names are listed in the BIOSCI Information sheet on the Web at http://www.bio.net/. For the METHODS-AND-REAGENTS list the USENET newsgroup name is bionet.molbio.methds-reagnts, thus the appropriate commands are sub bionet-news.bionet.molbio.methds-reagnts unsub bionet-news.bionet.molbio.methds-reagnts These commands are included in a message addressed to mxt@dl.ac.uk, NOT to the newsgroup mailing addresses. As usual, include the text in the body of the message as text on the Subject: line is ignored. To unsubscribe from all the lists at the UK node, use unsub bionet-news Please note that if the address in the list is different than the one in your mail message header, you will not be able to unsubscribe by this method. If you have problems, please mail biosci@daresbury.ac.uk. 4) The BIOSCI user address and research interest directory. ----------------------------------------------------------- Please take this opportunity to add your name, address, and research interest information to the BIOSCI User Address Database if you have not already done so. You can fill out the address form directly through our Web page at URL http://www.bio.net/adrform.html. The address database is reindexed nightly for WWW access (the URL is http://www.bio.net/). If you are not directly on the Internet but can reach it by e-mail, please use our waismail server to access the user directory. waismail use is described above. You can also request a user address form by e-mail from biosci-help@net.bio.net. Please check your database entry from time-to-time to see if your address information is still up-to-date. Because of our limited personnel resources, we ask that you resubmit a *complete* form to revise your entry; we only replace complete entries and do not have resources to edit old forms. Sincerely, Dave Kristofferson BIOSCI/bionet Manager biosci-help@net.bio.net From owner-biotechniques@net.bio.net Mon Jan 22 22:00:00 1996 Path: biosci!biosci!not-for-mail From: f.munkonge@ic.ac.uk (Dr F Munkonge) Newsgroups: bionet.journals.letters.biotechniques Subject: Fluorescent labeling of DNA Date: 23 Jan 1996 06:56:01 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 8 Sender: daemon@net.bio.net Approved: MMCCARTHY@BIOTECHNET.COM Distribution: world Message-ID: <4do7f0$4m4@mserv1.dl.ac.uk> NNTP-Posting-Host: net.bio.net Could anyone please enlighten me on the state of the art in labeling DNA with fluorescence molecules (especially FITC and TRITC). Leads on commercial sources would also be gratefully appreciated. Thanks Felix (f.munkonge.icl.ac.uk) From owner-biotechniques@net.bio.net Mon Jan 22 22:00:00 1996 Path: biosci!biosci!not-for-mail From: klynn@students.wisc.edu (Karyn Lynn) Newsgroups: bionet.journals.letters.biotechniques Subject: TRIzol and RDA Date: 23 Jan 1996 12:16:39 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 15 Sender: daemon@net.bio.net Approved: MMCCARTHY@BIOTECHNET.COM Distribution: world Message-ID: NNTP-Posting-Host: net.bio.net Can anyone comment on their results using TRIzol Reagent to isolate RNA or MesseageMaker kit for mRNA isolation (from Life Technologies)? How does it compare to the usual phenol/chloroform isolation protocols? I would like to isolate the RNA from Arabidopsis and use it to make cDNA and, subsequently, perform PCR on the cDNA. Also, is anyone successfully using representational difference analysis (RDA) on cDNA? I'd like to ask a few questions about the procedure. Karyn Lynn University of Wisconsin-Madison Genetics Department klynn@students.wisc.edu From owner-biotechniques@net.bio.net Tue Jan 30 22:00:00 1996 Path: biosci!biosci!not-for-mail From: mgl5403@centrum.dk (Med. Gas. Lab.) Newsgroups: bionet.journals.letters.biotechniques Subject: Re: TRIzol and RDA Date: 31 Jan 1996 07:58:01 -0800 Organization: Centrum Verden A/S Lines: 28 Sender: daemon@net.bio.net Approved: MMCCARTHY@BIOTECHNET.COM Distribution: world Message-ID: <4enftq$3cc@underworld.centrum.dk> References: NNTP-Posting-Host: net.bio.net klynn@students.wisc.edu (Karyn Lynn) wrote: >Can anyone comment on their results using TRIzol Reagent to isolate RNA or >MesseageMaker kit for mRNA isolation (from Life Technologies)? How does it >compare to the usual phenol/chloroform isolation protocols? I would like >to isolate the RNA from Arabidopsis and use it to make cDNA and, >subsequently, perform PCR on the cDNA. >Also, is anyone successfully using representational difference analysis >(RDA) on cDNA? I'd like to ask a few questions about the procedure. >Karyn Lynn >University of Wisconsin-Madison >Genetics Department >klynn@students.wisc.edu I have used TRIzol for isolation of RNA and subsequent RT-PCR. It works just fine. Easy protocol, but then the standard ones aren't that difficult. The reason for our use of TRIzol is that we have limited material (rectal biopsies) and want to do Western Blot on the same biopsy-material. Good luck on the project J. Hendel